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Appropriate irrelevant Abs were used as controls. Cells were fixed in paraformaldehyde and data were acquired http://www.selleckchem.com/products/MDV3100.html with FACSCalibur? or BD?LSR flow cytometers (BD Biosciences). Detection of IL-2, IL-10 and IFN-�� were performed by ELISA according to manufacturers�� instructions (BD Biosciences). GraphPad Prism (version 4.00 for Windows; GraphPad Software, San Diego, CA, USA) was used for statistical analyses. Results are expressed as mean �� SD, if one representative experiment is shown, or as mean �� SEM if data are averaged from more than one experiment. Comparison between two groups was performed by Student's t-test. Graft survivals were compared by Kaplan�CMeier analysis and the log-rank test. A p- value http://www.selleckchem.com/products/gsk1120212-jtp-74057.html we used BALB/c maturation-resistant (MR)-DCs generated in vitro (Figure S1B), which exhibit low allostimulatory potential and, when administered i.v., prolong cardiac allograft survival (Figure 1A). Injection of BALB/c MRDC in CD11c-DTR-B6 chimeras triggered division of 1H3.1 T cells specific for IE��52�C68 (BALB/c)-IAb (B6) (Figure S1C) only in not DC-depleted mice, indicating that cDCs are the only recipient professional APCs that present donor-Ag transferred from the injected MR-DCs since, in this model, plasmacytoid DCs are not depleted by DT injection (Figure S1D) (31,32). To evaluate the role of recipient DCs in ISDC-therapy in cardiac transplantation, the following key parameters were considered: (i) the optimal effect http://www.selleck.cn/products/pd-1-pd-l1-inhibitor-2.html is achieved when ISDCs are administered 7 days before transplantation (2�C4), (ii) recipient APCs in lymphoid tissues are necessary for elicitation of acute rejection (33�C35) and (iii) donor-derived ISDCs die within 3 days after i.v. administration (17). We confirmed in CD11c-DTR-B6 chimeras, that CD11chigh DCs are DT-depleted (Figure S2A) during the 3-day window when donor-Ag from the i.v.-injected ISDCs remains in the spleen (the latter assessed by proliferation of 1H3.1 T cells, Figure S2B). We also proved that after DT-administration, splenic CD11chigh DCs recover in number by the time of transplantation (day 0), so that they contribute to elicitation of rejection (Figure S2A). Administration BALB/c MR-DCs in CD11c-DTR-B6 chimeras 7 days before transplantation prolonged (p